Construction of chimeric human-rat IgH loci by BAC stitching and counterselection

Examples of targeting constructs for animal models and BAC modifications : Construction of chimeric human-rat IgH loci by BAC stitching and counterselection

 

Gene Bridges generated chimeric human-rat IgH loci using the Red/ET recombination technology for human antibody expression studies in transgenic rats. Therefore a 130 kb construct was assembled by combining DNA fragments of interest from three different BACs coding for the rat IgH locus by several Red/ET recombination steps (BAC stitching). Finally a part of the fused DNA was humanized by exchanging a 2 kb domain via a counter selection by Red/ET recombineering.

Voir tous les Kits de recombinaison Red/ET :
redet-kit
Scheme of BAC stitching and final humanizing of a 130 kb construct
chimeric

 

Ma B, Osborn MJ, Avis S, Ouisse LH, Mènoret S, Anegon I, Buelow R and Brüggemann M, 2013, Human antibody expression in transgenic rat: Comparison of chimeric IgH loci with human VH, D and JH but bearing different rat C-gene regions, J of Immunological Methods, 400-401, 78